MARCH 2013

Perceived and Potential Human

Health Risks Associated with

Consumption of Genetically

Modified Animals

Corinne Krentz1 Summary • The GM EnviropigTM produces an • Numerous varieties of genetically enzyme that enables phosphate modified (GM) crops have been absorption and utilization, thus reducing approved in Canada, and processed phosphate content of manure and foods containing GM ingredients can environmental effects associated with be found on grocery store shelves. In excess phosphate. No adverse effects contrast, there are currently no GM resulting from consumption of this animals or products derived from GM enzyme have been reported. animals approved for human consumption in Canada. • To date, there is little evidence that GM animals pose a significantly greater risk • According to the principle of to human health than their unmodified substantial equivalence, which is used counterparts in terms of allergenicity, to evaluate the safety of GM foods, anti-nutrient effects or acute toxicity. the genetically modified product must However, there is some uncertainty be at least as safe as its unmodified regarding long-term health effects. counterpart. Common types of health effects include protein allergenicity, anti-nutrient properties and toxicity. Introduction

• This evidence review summarizes the Genetically modified products have a growing potential health effects associated presence in the Canadian and international food with consumption of two GM animal market, with over 60 varieties of genetically products (GM Atlantic and altered plants approved for safe consumption in pork) that are currently undergoing 1 Canada. This does not include fruits and food safety review in North America. vegetables whose genetic composition has been • The GM AquAdvantage® Salmon modified using conventional methods such as produces excess growth hormone, breeding and selection. Organisms that are allowing it to reach market size more bioengineered to possess a particular trait rapidly. Due to the lack of animal through insertion of a specific and its feeding studies, long-term effects of required components for expression of a consuming GM are functional gene product are termed transgenic or unclear. genetically modified (GM).

1 Master of Public Health Program, University of Guelph 1

Often, the insertion of a particular gene conveys a This evidence review describes the elements required to desirable trait that may enhance the nutritional value of assess safety of genetically modified animals and the food, or give the transgenic variety a survival addresses the following questions: (1) What types of advantage compared to the unmodified species. human health risks are associated with consumption of Examples include enhanced growth rate; tolerance to a genetically modified foods, in general? (2) What are the particular herbicide, insect or virus; and ability to tolerate general attitudes among Canadian consumers toward a certain environmental condition (i.e., drought). Novel genetically modified animals intended for human cellular functions that convey desirable traits are consumption? (3) What are some of the health concerns achieved through insertion of a foreign gene (i.e., that associated with consumption of transgenic animals that originated from a different species), whereas are currently undergoing food safety review? enhancement of existing functions can be realized through insertion of additional copies of host . Methods Some crops have also been genetically modified to suppress undesirable traits, such as the production of The Web of Knowledge Database was used to search proteins that cause allergic reactions in humans. for relevant peer-reviewed articles. The search strategy used combinations of the key words [“transgenic” OR The process to make transgenic organisms involves: (1) “genetically modified” OR “GM”] AND [“health effects” creation of a construct, which is the genetic material to OR “human health” OR “safety”]. Articles referring to be inserted; (2) transfer of the construct into a single-cell genetically modified plants were excluded, as were fertilized egg; (3) incorporation of the genetic material those describing genetic modifications to animals for the into the host genome; and (4) selection for successful purpose of vaccine or drug production. Grey literature, incorporation of the construct into the host genome. such as consumer reports, government websites and Incorporation into the host genome enables the inserted GM animal safety assessment reports, was also used. gene to be passed down from generation to generation. Bacterial and viral vectors have been used in the creation of transgenic plants such as corn2,3 and rice.4 Assessment of Novel Food By comparison, most transgenic animals have been Safety created using to introduce construct DNA into the embryo.5,6 Opting not to use viruses or transposable elements (i.e., DNA that can cut and paste Principles of the Codex Alimentarius are used internationally to guide the risk assessment process for itself at different locations in the genome) in the creation 8 of the transgenic animal reduces the potential for novel foods. The process to evaluate the risk of GM unintended movement of genetic material within the food products is based on the concept of substantial genome, which is an important safety consideration if equivalence. That is, the genetically modified product the animal or its products are intended for human must be at least as safe as its unmodified counterpart, consumption. Another aspect to consider is public and it is desirable for the GM food to be at least as perception of risks associated with consumption of GM nutritious. Given the unique nature of each genetic animal food products. modification performed, the specific parameters required to evaluate the biological impacts of the new gene will While several transgenic animals intended for human vary on a case-by-case basis. consumption are in an advanced stage of development, to date none have received safety approval for market The risk assessment process requires an extensive sale from the United States Food and Drug Adminis- analysis of complex molecules that indicate nutritional tration (FDA) or Canadian regulatory agencies. In value (i.e., omega-3 fatty acids, trans fats), vitamins and Canada, regulatory approval for transgenic foods is minerals, and molecules with biological roles that may obtained through Health Canada, and these foods are be impacted by the newly introduced gene. A weight-of- subject to the Novel Foods Regulation, under Division evidence approach that considers the outcomes 28 of the Food and Drug Regulations.7 Applications for generated from all comparative tests is then used to approval of transgenic animals for food safety are determine whether there is a substantial difference in currently treated as “new animal drugs.” Two transgenic the risk of consuming the transgenic food. animals in the development pipeline are the EnviropigTM (Ontario Pork, ON) and AquAdvantageTM Salmon (AquaBounty Technologies, Inc., MA).

Potential Health Risks reactions. The nine foods that account for the vast majority of allergic reactions include peanuts, tree nuts, Associated with Genetically sesame seeds, eggs, milk, wheat, soybeans, seafood 13 Modified Foods and sulphites. If the newly inserted gene is derived from one of these foods, this should alert regulators that the transgenic food poses a greater risk for Proteins administered via the oral exposure route hypersensitive individuals compared to the non-GM generally pose a low risk because ingested proteins counterpart.14 This is illustrated in a case where a gene have low potential to be biologically active in the from the Brazil nut that encodes the protein 2S albumin bloodstream. Proteinaceous material in GM and non-GM was inserted into a soybean.15 The purpose of the foods undergoes degradation to amino acids in the modification was to improve nutritional quality; however, human stomach and intestine, and intact protein is one of the unintended effects of the transgenic soybean unlikely to enter the bloodstream since large molecules was its enhanced allergenic potential, affecting cannot be absorbed by the gut. High temperatures from individuals with hypersensitivity to Brazil nuts.15 cooking also contribute to protein denaturation. The second step may involve comparative analysis to One study that seemingly contradicted this paradigm determine amino acid sequence and/or structural reported that recombinant protein produced in GM similarity between the novel protein and proteins that are 9 maize was found in human blood. However, one of the known allergens.16 In vitro testing of purified protein main limitations of this study is that the commercial (encoded by the gene of interest) using immunoassays, antibody-based assay used to quantify the protein is and in vivo testing to assess tolerance using animal intended for use to analyze plant seeds and leaves, and models, will also indicate allergenicity potential of the 10 has been shown to perform poorly for blood analyses. protein. Animal models that are considered appropriate There may be proteins in blood that cause a false- for assessment of allergenicity include mice and rats positive test result and the study did not contain (i.e., Brown Norway rat, BALB/c mouse), dogs and negative controls (i.e., serum from individuals who had pigs.14 Proteins evaluated using in vivo models may be not been exposed to the recombinant protein) to administered orally or by subcutaneous injection to investigate this possibility. Furthermore, other studies achieve greater sensitivity. Allergenicity studies investigating the fate of the same protein (Cry1Ab) in conducted using piglets are considered a very good cattle that were fed GM corn did not detect fragments of approximation of the response that can be expected in 11 the protein in blood, milk or urine samples. humans.14

Adverse human health effects associated with Lastly, skin-prick tests using human subjects can genetically modified foods can be categorized as (1) provide an indication of whether the protein raises an allergenicity to the protein encoded by the newly IgE-mediated inflammatory response. Transgenic food introduced gene, (2) anti-nutrient effects and (3) toxicity. containing proteins found to be allergens would require that it be labelled accordingly to alert consumers. Allergenicity Anti-Nutrient Effects Approximately 4% of Canadians have a food allergy.12 Proteins in food products can cause an acute In some cases, a newly introduced gene can encode a inflammatory response of the skin, mouth or digestive protein that negatively affects the uptake and/or tract in some individuals. These reactions can range utilization of a particular nutrient. For example, from mild inflammation to severe anaphylactic shock. compounds found in soybeans decrease the enzymatic The potential for the protein product of the newly activity of trypsin, thereby interfering with protein 17 introduced gene to cause an allergic reaction is a key digestion and uptake of essential amino acids. Each safety consideration. food has a unique profile of nutrients, anti-nutrients and toxins. All of these key compounds require analysis in An approach to evaluate allergenicity was developed by both the GM and the unmodified counterpart to the International Food Council and the determine the impacts of the introduced gene on food International Life Sciences Allergy and Immunology composition. Institute. The principles of this approach involve first determining whether the gene is derived from, or also An additional consideration is the impacts of food present in, an organism known to cause hypersensitivity processing on nutrients, anti-nutrients and toxins, as these may alter levels of available nutrients or create 3 undesirable compounds in the finished product. mortality rates and tumour incidence are greater than Proteinaceous anti-nutrients denature at high would be expected due to chance. temperatures, which may affect biological function and nutrient availability. Fermentation may reduce levels of Another issue is whether toxic effects may result from certain compounds such as phytic acid, which chelates consumption of herbicide-resistant plants that contain with zinc and other minerals.18 Thus, fermentation can residual chemical herbicide. Residues of herbicides increase the bioavailable levels of some minerals. used on GM crops have been detected in human blood, but the levels are well below those that are considered For transgenic animals intended for human harmful.10 consumption, this analysis should be conducted for both raw and cooked sections of all parts of the animal that Perceived Risks and Attitudes may be consumed. about GM Animals Toxicity Studies have indicated that consumer acceptance of GM No reports of acute toxic effects in humans or animals foods varies in different countries.24,25,26,27 The success as a result of consuming GM plants have been reported. of any GM product is highly dependent upon consumer Toxicity studies are usually conducted for a 90-day preferences, which are shaped in part by attitudes about period to assess acute health effects. While some genetically modified organisms and the perceived risks differences between groups of animals fed GM and non- and benefits of the product. GM diets have been observed in this period,19,20 the physiological significance of these observations is While GM foods can provide benefits to the consumer unclear. Long-term studies where subjects are fed GM including enhanced nutritional value, price advantage, diets for at least two years are required to provide improved shelf life and better taste, these benefits may evidence that the changes observed in the first 90 days be insufficient to outweigh the consumer’s perception of are signs of toxicity that will progress to chronic disease, risks about GM products. Negative attitudes about GM and to adequately monitor tumour incidence and food can be related to (1) concerns about perceived progression. human health risks; (2) a lack of trust in institutions responsible for ensuring food safety and (3) unintended Several long-term feeding studies have been conducted environmental consequences of GM food production to assess the safety of animal feed containing GM corn such as resistance, gene transfer and threats that produces Bacillus thuringiensis or Bt toxin, which is to biodiversity such as through interbreeding between lethal to some types of insects. Dairy cows fed GM corn GM and non-GM animals. for 25 months had slightly lower body weights than 21 those in the control group. Diet had no effect on milk GM produce and processed foods containing GM yield, but minor differences in milk composition were ingredients can be found on grocery store shelves observed. Flachowsky et al (2005) found no meaningful around the world, but transgenic meat products have not differences between quails fed a diet supplemented with yet received market approval in any country to date. 22 GM corn and quails fed an isogenic variety of corn. Consumers are generally less accepting of genetic modifications to animals compared to plants.24 In A recent life-long study of rats was conducted to Canada, transgenic animal products intended for human evaluate long-term effects associated with consumption consumption are a subject of controversy. Anti-GM 23 of herbicide-resistant corn (NK603). It included a group messages from public interest groups and organizations of rats fed a diet supplemented with Roundup-tolerant such as the Canadian Biotechnology Action Network corn that had not been sprayed with Roundup herbicide. and negative publicity from media, describing GM One limitation of this study was the small number of rats salmon as “Frankenfish,”28,29 have contributed to the (10) used per gender-controlled treatment group. A high stigmatization of GM animal products. proportion of control and GM-fed female rats developed mammary tumours. Mortality frequency of the male A survey of 1,300 Canadians in 2005 indicated that control group was slightly greater than that of male GM- many consumers had negative predispositions towards fed groups at some dietary doses, while mortality transgenic animals.30 Some of the concerns about frequency of the female control group was lower than transgenic animals intended for human consumption that of all female GM-fed groups. Replication of this that were expressed by survey respondents included study is required to determine whether differences in potential side effects resulting from the increased

4 hormone levels, unknown long-term consequences and weight are GM.32 In comparison, Europe, Australia and 33 increased risk of cancer. Further, the study found that Japan have a mandatory GM food labelling policy. disclosure of product risks and benefits did not have a Products that contain a percentage of GM ingredients negative effect on their perceptions of transgenic animal that is greater than the threshold prescribed by national products or intent to purchase them in the future. In regulators must have a GM label. In Europe and order for consumers to make informed decisions about Australia, foods require a label if more than 1% of any the products they purchase, it is paramount for ingredient by weight is genetically modified. manufacturers to not only clearly communicate the benefits of GM products, but also to label GM products GM Animals in Advanced Stages accordingly. of Development It is not uncommon for processed foods in North America to contain genetically modified ingredients, GM animals and GM animal-derived products intended since many corn and soybean crops are transgenic for human consumption that are currently under varieties. A consumer report’s study conducted in 1999 development, but not yet approved for market sale by confirmed that genetically modified ingredients from regulatory bodies, are summarized in Table 1. The risks crops were present in grocery store foods such as infant and benefits associated with two GM animals intended 31 formulas, veggie burgers, tortilla chips and muffin mix. for human consumption, the AquAdvantageTM Salmon In North America, these products do not require labels and the EnviropigTM, are discussed in more detail. indicating that they contain GM ingredients.

Canada and the United States have adopted a voluntary labelling policy. Manufacturers are not required to label foods containing GM ingredients. Foods can be labelled “GM-free” if less than 5% of the total ingredients by

Table 1. Transgenic animals and animal-derived products intended for human consumption that are in development

Protein encoded Transgenic animal Gene source Benefit Reference by new gene Animals intended for human consumption Yorkshire Pig Phytase Escherichia coli Enables utilization of [34] (EnviropigTM) phosphorus in phytic acid Reduces need to feed pigs phosphorus supplements Reduced phosphorus in manure and environment Pig Growth hormone Porcine Enhanced growth rate [35] Pig Humanized n-3 fatty Caenorhabditis Enhanced nutritional [36] acid desaturase elegans value by increasing omega-3 fatty acid content Atlantic salmon Growth hormone Enhanced growth rate [37] (AquAdvantageTM) Amago salmon Growth hormone Enhanced growth rate [38] Chicken Recombinant Mx Mouse Enhanced ability for [38] chickens to resist viral infection Cow Mammalianized n-3 Caenorhabditis Enhanced nutritional [40] fatty acid desaturase elegans value by increasing omega-3 fatty acid content

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Protein encoded Transgenic animal Gene source Benefit Reference by new gene Animal-derived products intended for human consumption Goat Lysozyme Human Enhanced antimicrobial [41] properties in milk Cow Recombinant Human Enhanced antimicrobial [42] Lysozyme properties in milk Cow β-casein, Bovine Enhanced nutritional [43] κ-casein value and characteristics for processing

AquAdvantageTM Salmon elevated levels of hormones. The hormones responsible for accelerated growth in the transgenic In the 1990s, a transgenic Atlantic salmon was fish are GH and insulin-like growth factor-I (IGF-I), developed that differed from its unmodified and it is unknown whether the amounts present in the counterpart in two keys ways. Firstly, the transgenic fish are sufficient to cause undesirable long-term salmon contained genetic material that encodes a effects in humans. IGF-I is a signaling molecule that Chinook salmon growth hormone. Expression of the plays a key role in bone and muscle growth. At the gene encoding the growth hormone (GH) enabled cellular level, it promotes cell proliferation and fish to grow at an accelerated rate, up to six times differentiation. GH acts primarily through stimulation 37 greater than that of non-transgenic fish. The of liver cells to produce IGF-I. Levels of GH are increase in growth rate reduced the time required for controlled by a negative feedback system where high the fish to be harvested. Secondly, the transgenic levels of IGF-I cause suppression of GH, and thereby salmon were engineered to be female and triploid IGF-I. Thus, the biological effects of consuming food (i.e., have three sets of chromosomes), rendering containing elevated levels of GH and IGF-I require them incapable of successfully reproducing. Thus, in investigation. the unlikely event that these salmon were released into the wild, they would pose no long-term threat to The potential for GH and IGF-I in fish tissue to exert native fish species. effects on humans who consume the fish depends on several factors including level of exposure (i.e., how The transgenic salmon was named the much fish is consumed), duration of exposure (i.e., TM Salmon and is being taken to AquAdvantage how frequently fish are consumed), and how much of market by AquaBounty Technologies, Inc. Proposals the functional protein is absorbed and reaches the to the FDA and Health Canada for food safety target site before being degraded by proteases. approval have been under review since 2010. The Despite the fact that proteins administered orally are document submitted to the FDA contained likely to undergo degradation from digestive comparative data from the transgenic and wild enzymes, no animal feeding studies have been Atlantic salmon to evaluate potential human health conducted to evaluate the effects of consuming risks associated with the transgenic variety.44 The transgenic fish containing elevated levels of GH. conclusions in the report stated that no biologically Such studies should be required as part of a rigorous relevant differences between transgenic TM investigation of potential health effects, especially for AquAdvantage Salmon and its non-transgenic compounds such as hormones. counterpart were found, and that AquAdvantageTM Salmon posed no threat to the environment. Few studies have compared concentrations of growth Recently the FDA also concluded that transgenic hormone in plasma/tissues of transgenic and non-GM salmon would have no significant environmental Atlantic salmon. Studies showing no significant impact in the United States. However, the report difference between plasma concentrations of growth lacked discussion on potential short- and long-term hormone in GM verses non-GM species had human health impacts. inadequate sample sizes (i.e., 5 to 7 fish per group).37,44 Concentrations of growth hormone in tissues from GM and non-GM salmon have been Potential Human Health Effects Related to 44 Consumption of AquAdvantageTM Salmon below the quantification limit of the method used.

One major human health concern with Another issue is the species-specificity of fish GH AquAdvantageTM Salmon relates to the uncertainty and whether fish GH has biological activity in associated with consumption of a food that contains humans. Fish and human GH proteins are 6 biologically different, sharing only 33% amino acid non-transgenic pigs, and the amount of land required sequence similarity. No in vitro or in vivo studies to to spread manure is reduced by approximately one- evaluate biological activity of fish GH on human GH third.34 receptors have been conducted. However, early fish transgenic experiments performed using human GH Pigs are typically fed phosphorus supplements to in fish showed that transgenic fish grew more rapidly meet their dietary need, since pigs are unable to and were larger than non-GM fish, indicating that digest phytic acid, the storage form of phosphorus in cross-species interactions between human GH and 45 grains. Thus, pig feces contain high levels of fish GH-receptors may occur. indigestible phosphorus. Alternatives to phosphorus supplements include feeding the animals digestive High levels of IGF-I and GH have been found to be enzymes or diets that are high in digestible forms of associated with higher risk of developing some types phosphorus and low in phytic acid, or genetically of cancers and increased aggression of existing engineering pigs to produce a greater repertoire of 46 tumours in humans. Acromegalic patients having digestive enzymes. excess secretion of GH from the pituitary gland were TM shown to have a higher prevalence of risk factors Enviropigs were developed using microinjection (i.e., hypertension and diabetes) for cardiovascular technology to express a gene encoding the enzyme disease, which is the leading cause of their phytase, which is not normally produced by 47 mammals. Phytase catalyzes the hydrolysis of phytic mortality. Heart abnormalities were observed acid into inorganic phosphorus that can be absorbed among some of the GM salmon in previous studies, in the small intestine. Phytic acid has anti-nutrient indicating that this area of research may warrant 44 properties that are remedied by phytase. Phytic acid further attention. is a negatively-charged molecule that chelates with monovalent (i.e., K+) and divalent cations (i.e., Ca2+, A sequence similarity assessment between salmon Zn2+, Mg2+), making these nutrients unavailable for GH and a database of allergens was performed to absorption in the intestine. Phytases are produced 44 assess allergenicity potential. Salmon GH did not naturally by a wide range of bacteria, fungi and share amino acid sequence homology with any of the plants.48 The phytase enzyme from Escherichia coli allergens in the database. Homology was defined as was used to create the EnviropigTM. Genes encoding having a minimum of 35% identity in a contiguous phytase enzymes have also been used in the sequence of 80 amino acids. Although in vitro testing creation of transgenic rice, wheat, sugarcane, was not performed to compare the levels of proteins soybean, canola and potato to enhance nutrient and that are known allergens in GM and non-GM Atlantic mineral bioavailability.49 salmon, a similar transgenic salmon was evaluated in vitro using proteins from GM and non-GM fish (n=3 Since the initial development of the EnviropigTM, other per group), and in vivo using sera from patients with technologies have been developed to address the fish allergies.40 They found that levels of known fish issue of excess nutrients in livestock manure. These allergens were not significantly different in GM include processing slurries to extract phosphate in a compared to non-GM fish. crystallized form (struvite), which can be used as a fertilizer.50 Alternatively, feeding pigs digestive No anti-nutrients or toxic substances in the enzymes has been shown to reduce phosphorus in transgenic salmon have been identified. While it may feces by 30 to 40%.51,52 be unlikely that consumption of AquAdvantage TM Salmon has unforeseen anti-nutrient or toxic effects, Ontario Pork was a great supporter of EnviropigTM no animal feeding studies using diets supplemented research and development for several years, and TM with AquAdvantage Salmon have been conducted, holds the trademark, but stopped backing the to our knowledge, to confirm this. transgenic pig in 2012. Commercialization TM TM opportunities for the Enviropig are being explored. Enviropig TM Potential Human Health Effects Related to The Enviropig was developed in the 1990s by TM scientists at the University of Guelph to address a Consumption of Enviropig pressing environmental issue. Phosphorus is often a Some scientists argue that phytase has a range of growth-limiting nutrient in aquatic environments and health benefits in humans.49 However, these human agricultural run-off containing high levels of health benefits would be the result of consuming raw phosphorus can harm aquatic ecosystems. foods that naturally contain phytase. Phytase protein Phosphorus-rich inputs to lakes and rivers can cause denatures when it is heated, thus cooked pork would oxygen depletion, fish kills and/or algal blooms. not be a significant source of enzymatically active EnviropigsTM excrete up to 60% less phosphorus than phytase.

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In terms of allergen potential, phytase in powder form regulatory body in the world, and many consumers can cause respiratory problems such as asthma are not convinced that transgenic food products, 53,54 when inhaled in an occupational setting. especially those derived from animals, are completely However, no adverse immunological effects as a safe. The safety of novel foods is currently assessed result of phytase consumption have been reported. using the concept of substantial equivalence, where No toxic effects resulting from phytase consumption potential risks of the transgenic and non-transgenic product are compared. To date, evidence indicates have been reported in animal feeding studies. One TM TM feeding study by Zhang et al. (2000) compared the that AquAdvantage Salmon and Enviropig are not health effects of administering different forms of different from their unmodified counterparts in terms phytase to pigs.52 One group of pigs was given of their nutritional value, allergenicity, toxicity and phytase in the form of a commercial feed enzyme anti-nutrient effects. One limitation, however, is that and members of the other group were fed transgenic long-term risks have not been thoroughly assessed canola that contained a gene encoding phytase. and additional animal feeding trials are needed. Histological and macroscopic analysis of organ Genetically modified animal products will likely tissues of pigs that were fed diets supplemented with remain off grocery store shelves until the uncertainty phytase in either form did not reveal any toxic surrounding potential long-term health effects is effects.52 reduced. Looking forward, the benefits of GM animal products need to be clearly communicated to consumers if the GM product is to succeed in today’s Knowledge Gaps competitive food market.

• Additional research on potential long-term effects of consuming genetically modified Acknowledgements foods is required. These long-term studies (two years or more) will shed light on whether We would like to thank Helen Ward, Sophie Verhille, some of the metabolic differences observed Mike Brauer, and Robert Friendship for their in short-term (90 day) animal feeding studies invaluable input and review of this document. are pathologically relevant and will monitor effects that occur at later stages such as tumours. References • Animal feeding trials to study the effects of consuming cooked transgenic fish or pork 1. Health Canada. 2011. Genetically modified foods are needed. and other novel foods. Date modified 2011-03-07. http://www.hc-sc.gc.ca/fn-an/gmf-agm/appro/index- • Additional experiments with adequate sample eng.php sizes are needed to verify that there is no 2. Choi I R, Stenger D C, Morris T J, French R. A plant biologically significant difference between virus vector for systemic expression of foreign genes in levels of hormones such as GH and IGF-I in cereals. The Plant Journal. 2000; 23(4): 547-555. transgenic and non-transgenic salmon tissues. 3. Negrotto D, Jolley M, Beer S, Wenck A R, Hansen G. The use of phosphomannose-isomerase as a selectable marker to recover transgenic maize plants • The most recent consumer opinion survey in (Zea mays L.) via transformation. Plant Canada on attitudes toward transgenic Cell Rep. 2000; 19(8): 798-803. animals was conducted in 2005. It would be interesting to re-evaluate consumer attitudes 4. Hiei Y, Komari T, Kubo T. Transformation of rice in light of new information that has since mediated by agrobacterium tumefaciens. Plant Mol emerged regarding transgenic organisms. Biol.1997; 35(1-2): 205-218. 5. Hammer R E, Pursel V G, Rexroad C E, Wall R J, Bolt D J, Ebert K M, Palmiter R D, Brinster R L. Conclusions Production of transgenic rabbits, sheep and pigs by microinjection. Nature. 1985; 315(20): 680-683. TM TM The AquAdvantage Salmon and Enviropig are 6. Bleck G T, White B R, Miller D J, Wheeler M B. among the first genetically modified animal products Production of bovine α-lactalbumin in the milk of that were developed to address food security and transgenic pigs. J Anim Sci. 1998; 76(12): 3072-3078. environmental issues. These transgenic animals 7. Department of Justice. 2012. Food and drug have yet to receive safety approval from any regulations. http://laws-

8 lois.justice.gc.ca/eng/regulations/C.R.C.%2C_c._870/pa 19. Séralini G E, Cellier D, de Vendômois J S. New ge-191.html#h-144 analysis of a rat feeding study with a genetically modified maize reveals signs of hepatorenal toxicity. 8. World Health Organization. Codex Alimentarius. Arch Environ Contam Toxicol. 2007; 52(4):596-602. International food standards. 2012. http://www.codexalimentarius.org/standards/list-of- 20. de Vendômois J S, Roullier F, Cellier D, Séralini G standards/en/ E. A comparison of the effects of three GM corn varieties on mammalian health. Int J Biol Sci. 2009; 9. Aris A, Leblanc S. Maternal and fetal exposure to 5(7):706-726. pesticides associated to genetically modified foods in Eastern Townships of Quebec, Canada. Reprod 21. Steinke K, Guertler P, Paul V, Wiedemann S, Ettle Toxicol. 2011; 31(4): 528-533. T, Albrecht C, Meyer H H D, Spiekers H, Schwarz F J. Effects of long-term feeding of genetically modified corn 10. Chowdhury E H, Kuribara H, Hino A, Sultana P, (event MON810) on the performance of lactating dairy Mikami O, Shimada N, Guruge K S, Saito M, Nakajima cows. J Anim Physiol Anim Nutrition. 2010; 94(5): e185- Y. Detection of corn intrinsic and recombinant DNA e193. fragments and Cry1Ab protein in the gastrointestinal contents of pigs fed genetically modified corn Bt11. J 22. Flachowsky G, Halle I, Aulrich K. Long term feeding Anim Sci. 2003; 81(10): 2546-2551. of Bt-corn – a ten-generation study with quails. Arch Anim Nutrition. 2005; 59(6): 449-451. 11. Guertler P, Vijay P, Steinke K, Wiedemann S, Preissinger W, Albrecht C, Spiekers H, Schwarz F J, 23. Séralini G E, Clair E, Mesnage R, Gress S, Defarge Meyer H H D. Long-term feeding of genetically modified N, Malatesta M, Hennequin D, de Vendômois J S. Long corn (MON810) – fate of cry1Ab DNA and recombinant term toxicity of a Roundup herbicide and a Roundup- protein during the metabolism of the dairy cow. tolerant genetically modified maize. Food Chem Toxicol. Livestock Sci. 2010; 131(2-3): 250-259. 2012; 50(11); 4221-4231. 12. Ben-Shoshan M, Harrington D, Fragapane J, Soller 24. Nayga R M, Fisher M G, Onyango B. Acceptance of L, Joseph L, Pierre Y, Godefroy S, Elliott S, Clarke A. genetically modified food: comparing consumer Food allergies in Canada: prevalence and associated perspectives in the United States and South Korea. Agr factors. J Allergy Clin Immunol. 2010; 125(2): 215. Econ. 2006; 34(3): 331-341. 13. Canadian Food Inspection Agency. Food allergies 25. Costa-Font M, Gil J M. Structural equation modelling and allergen labelling information for consumers. of consumer acceptance of genetically modified (GM) Ottawa, On: CFIA. food in the Mediterranean Europe: a cross country http://www.inspection.gc.ca/food/consumer-centre/food- study. Food Qual Prefer. 2009; 20(6): 399-409. safety-tips/labelling-food-packaging-and- 26. Kikulwe E M, Wesseler J, Falck-Zepeda J. Attitudes, storage/allergen/eng/1332442914456/1332442980290 perceptions, and trust. Insights from a consumer survey 14. Ahuja V, Quatchadze M, Ahuja V, Stelter D, Albrecht regarding genetically modified banana in Uganda. A, Stahlmann R. Evaluation of biotechnology-derived Appetite. 2011; 57(2): 401-413. novel proteins for the risk of food-allergic potential: 27. Mather D W, Knight J G, Insch A, Holdsworth D K, advances in the development of animal models and Ermen D F, Breitbarth T. Social stigma and consumer future challenges. Arch Toxicol. 2010; 84(12): 909-917. benefits: trade-offs in adoption of genetically modified 15. Nordlee J A, Taylor S L, Townsend J A, Thomas L foods. Sci Comm. 2012; 34(4): 487-519. A, Bush MD. Identification of a Brazil-nut allergen in 28. Blouin D. Frankenfish: just another monster? transgenic soybeans. New Engl J Med. 1996; Canadian Geographic. 2004; Sept/Oct. 334(2):688-692. http://www.canadiangeographic.ca/magazine/so04/inde 16. Zehong Z O, Ying H E, Lin R, Baoqing S U, Huifang pth/ C, De C, ShiMing L, XiaoGuang Y, AiLin T. A 29. Walsh B. Frankenfish: is GM salmon a vital part of bioinformatic evaluation of potential allergenicity of 85 our future? Time Science. 2011. candidate genes in transgenic organisms. Chinese Sci http://www.time.com/time/health/article/0,8599,2082630, Bull. 2012; 57(15):1824-1832. 00.html 17. Organization for Economic Co-operation and 30. Castle D, Finlay K, Clark S. Proactive consumer Development. Consensus document on compositional consultation: the effect of information provision on considerations for new varieties of soybean: key food response to transgenic animals. J Public Affairs. 2005; and feed nutrients and anti-nutrients. Paris: OECD, 5: 200-216. 2001. http://www.oecd.org/dataoecd/15/60/46815135.pdf 31. Consumer Reports. Seeds of change. September 1999; 64(9): 41. 18. Liener I E. Implications of antinutritional components in soybean foods. Critical Reviews in food Science and 32. Gruère G. A review of international labeling policies Nutrition. 1994; 34(1):31-67. of genetically modified food to evaluate India’s proposed rule. AgBioForum. 2007: 10(1): 51-64.

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33.Gruère G, Carter C, Farzin Y. What labelling policy 44. Food and Drug Administration. Center for Veterinary for consumer choice? The case of genetically modified Medicine. AquAdvantage salmon. 2010. food in Canada and Europe. Can J Econ. 2008; 41(4): http://www.fda.gov/downloads/AdvisoryCommittees/Co 1472-1497. mmitteesMeetingMaterials/VeterinaryMedicineAdvisory Committee/UCM224762.pdf 34. Forsberg C W, Phillips J P, Golovan S P, Fan M Z, Meidinger R G, Ajakaiye A, Hilborn D, Hacker R R. The 45. Chen T T, Lin C M, Zhu Z, Gonzalez-Villasenor L I, Enviropig physiology, performance, and contribution to Dunham R A, Powers D A. Gene transfer, expression nutrient management advances in a regulated and inheritance of and human growth environment: the leading edge of change in the pork hormone genes in carp and loach. UCLA Symposia on industry. J Anim Sci 2003; 81: E68-E77. Molecular and Cellular 1990; 116: 127-139. 35. Vize P D, Michalska A E, Ashman R, Lloyd B, Stone 46. Kauppinen-Mäkelin R, Sane T, Välimäki M J, B A, Quinn P, Wells J R E, Seamark R F. Introduction of Markkanen H, Niskanen L, Ebeling T, Jaatinen P, a porcine growth hormone fusion gene into transgenic Juonala M, the Finnish Acromegaly Study Group, pigs promotes growth. J Cell Science. 1988; 90: 295- Pukkala E. Increased cancer incidence in acromegaly— 300. a nationwide survey. Clin Endocrinol. 2010; 72(2): 278- 279. 36. Lai L, Kang J X, Li R, Wang J, Witt W T, Yong H Y, Hao Y, Wax D M, Murphy C N, Rieke A, Samuel M, 47. Berg C, Petersenn S, Lahner H, Herrmann B L, Linville M L, Korte S W, Evans R W, Starzl T E, Prather Buchfelder M, Droste M. Cardiovascular risk factors in R S, Dai Y. Generation of cloned transgenic pigs rich in patients with uncontrolled and long-term acromegaly: omega-3 fatty acids. Nature Biotechnol. 2006; 24(4): population and the effect of disease control clinical 435-436. endocrinology. J Clin Endocrinol Metab. 2010; 95(8): 3648 -3656. 37. Du S J, Gong Z, Fletcher G L, Shears M A, King M J, Idler D R, Hew C L. Growth enhancement in 48. Rao D E C S, Rao K V, Reddy T P, Reddy V D. transgenic Atlantic salmon by the use of an “all fish” Molecular characterization, physicochemical properties, chimeric growth hormone gene construct. Nat known and potential applications of phytases: an Biotechnol. 1992; 10(2): 176-181. overview. Crit Rev Biotechnol. 2009; 29(2): 182-198. 38. Nakamura R, Satoh R, Nakajima Y, Kawasaki N, 49. Kumar V, Sinha A K, Makkar H P S, Becker K. Yamaguchi T, Sawada J, Nagoya H, Teshima R. Dietary roles of phytate and phytase in human nutrition: Comparative study of GH-transgenic and non- a review. Food Chem. 2010; 120(4): 945-959. transgenic amago salmon ( masou 50. Qureshi A, Lo K V, Mavinic D S, Liao P H, Koch F, ishikawae) allergenicity and proteomic analysis of Kelly H. Dairy manure treatment, digestion and nutrient amago salmon allergens. Regul Toxicol Pharm. 2009; recovery as a phosphate fertilizer. J Environ Sci Health. 55(3): 300-308. 2006; 41(7): 1221-1235. 39. Min S, Qing S Q, Hui Y Y, Zhi F D, Rong Q Y, Feng 51. Pfeiffer A. The influence of phytase in phosphorus- X, Chun L B. Generation of antiviral transgenic chicken reduced diets on the amount of P-excretion in the case using spermatogonial transfected in vivo. Afr J of fattening pigs and estimations of the amount of P- Biotechnol. 2011; 10(70): 15678-15683. excretion based on P-concentration in feces 40. Wu X, Ouyang H, Duan B, Pang D, Zhang L, Yuan respectively. Zuchtungskunde. 1995; 67(2): 147-157. T, Xue L, Ni D, Cheng L, Dong S, Wei Z, Li L, Yu M, 52. Zhang Z B, Kornegay E T, Radcliffe J S, Wilson J H, Sun Q-Y, Chen D-Y, Lai L, Dai Y, Li G-P. Production of Veit H P. Comparison of phytase from genetically cloned transgenic cow expressing omega-3 fatty acids. engineered Aspergillus and canola in weanling pig diets. Transgenic Res. 2012; 21(3): 537-543. J Anim Sci. 2000; 78(11): 2868-2878. 41. Maga E A, Jackson K, Archer G, Mench J A, Van 53. Bauer X. Enzymes as occupational and Eenennaam A L, Murray J D. Assessment of the well- environmental respiratory sensitizers. Int Arch Occup being and behavior of genetically engineered dairy Environ Health. 2005: 78(4): 279-286. goats. Trangenic Res. 2010; 19:137. 54. van Heemst R C, Sander I, Rooyackers J, de Jong 42. Yang B, Wang J, Guo C, Yu T, Sui S, Tang B, Li R, Liu Y, Dai Y, Zhou Q, Li N. Characterization of L, Djamin R S, Aerts J G, Belderbos H N A. recombinant human lysozyme expressed in the milk of Hypersensitivity pneumonitis caused by occupational cloned transgenic cows. Trangenic Res. 2010; exposure to phytase. Eur Respir. 2009; 33(6): 1507- 19(1):147. 1509. 43. Brophy B, Smolenski G, Wheeler T, Wells D, Huillier P L, Laible G. Cloned transgenic cattle produce milk with higher levels of β-casein and κ-casein. Nature Biotechnol. 2003; 21: 157-162.

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